Highly Efficient JFH1‐Based Cell‐Culture System for Hepatitis C Virus Genotype 5a: Failure of Homologous Neutralizing‐Antibody Treatment to Control Infection

Tanja B. Jensen, Judith M. Gottwein, Troels K. H. Scheel, Anne M. Hoegh, Jesper Eugen‐Olsen, Jens Bukh
2008 Journal of Infectious Diseases  
Background. Recently, a hepatitis C virus (HCV) cell-culture system was developed that employed strain JFH1 (genotype 2a), and JFH1-based intra-and intergenotypic recombinants now permit functional studies of the structural genes (Core, E1, and E2), p7, and NS2 of genotypes 1-4. The goal was to adapt the system to employ genotype 5. Methods. Huh7.5 cells infected with SA13/JFH1, containing Core-NS2 of strain SA13 (genotype 5a), were monitored for Core expression and for supernatant infectivity
more » ... nd HCV-RNA titers. Adaptive mutations of SA13/JFH1 were identified by sequence analysis of recovered genomes and reverse-genetic studies. Receptor blockage was performed with anti-CD81 and anti-SR-BI. For neutralization experiments, SA13/JFH1 or JFH1-based viruses of other genotypes were incubated with patient sera. Results. SA13/JFH1 with NS2 and NS3 mutations yielded infectivity titers Ͼ10 5 TCID 50 /mL. Infection with SA13/JFH1 was inhibited by CD81 blocking and SR-BI blocking, respectively, and by preincubation with genotype 5a chronic-phase patient sera. Such sera had varying cross-genotype neutralization potential. However, preincubation and treatment with homologous neutralizing antibodies could not control SA13/JFH1 infection in culture. Conclusion. The SA13/JFH1 culture permits genotype 5a-specific studies of Core-NS2 function and interfering agents. The ability of HCV to spread in vivo during treatment with neutralizing antibodies was confirmed in vitro.
doi:10.1086/593021 pmid:19032070 fatcat:bfzbirgqa5ewzoyfx576q52ojq